The Use of Regional Gene Delivery to Heal Critical Sized Bone Defects

使用区域基因传递来治愈严重骨缺损

• 批准号: 8055046
• 负责人: JAY R. LIEBERMAN
• 金额: $ 29.89万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8055046
• 关键词: Address; Adenovirus Vector; Adopted; Adoption; Arthroplasty; Asses; Biodistribution; Biological; Biological Assay; Biology; Bone Marrow; Bone Marrow Cells; Bone Morphogenetic Proteins; Bone Regeneration; Cell Differentiation process; Cells; Cherry - dietary; Cicatrix; Clinical; Clinical Data; Complementary DNA; Data; Defect; Development; Evaluation; Fluorescence; Fracture; Gene Delivery; Goals; Harvest; Healed; Human; Immune response; Implant; Joints; Knowledge; Laboratories; Lentivirus Vector; Life; Marrow; Methods; Modeling; Mus; One-Step dentin bonding system; Organ; Osteoblasts; Osteogenesis; Patients; Pericytes; Procedures; Process; Rattus; Recombinants; Reporter; Reporter Genes; Research; Research Proposals; Rodent; Role; Safety; Site; Smooth Muscle Myocytes; Solutions; Spinal Fusion; Surgeon; Technology; Time; Tissues; Toxic effect; Transgenic Mice; Transgenic Organisms; Treatment Protocols; Vascular blood supply; autocrine; bench to bedside; bone; bone cell; bone healing; bone loss; bone morphogenetic protein 2; bone quality; cellular transduction; comparative efficacy; cost effective; effective therapy; gene therapy; healing; implantation; interest; pre-clinical; progenitor; programs; promoter; public health relevance; red fluorescent protein; research study; response; treatment strategy; user-friendly; vector

DESCRIPTION (provided by applicant): Ex vivo regional gene therapy has the potential to solve difficult bone loss problems associated with large bone stock deficiency, abundant scar tissue and limited vascular supply for which at the present time there is no consistently satisfactory solution. We have demonstrated proof of concept the potential clinical utility of this strategey by successfully healing critical sized rodent femoral defects with rat and human bone cells transduced with either an adenoviral vector (Ad- BMP-2) or lentiviral vector (LV-BMP-2) containing the cDNA for BMP-2. In a recent comparison of healing of a bone defect with Ad-BMP-2 versus LV-BMP-2 better quality bone was obtained with the LV-BMP-2 treatment. We hypothesized that the more prolonged BMP expression associated with the lentiviral vector was responsible for the better results with the LV vector and that this increased duration of expression may be necessary to achieve adequate bone repair in the challenging biological enviorments seen in patients. In order for gene therapy to be adopted for clinical use it must be cost-effective, safe and there must be a better understanding of the biology of bone repair with transduced cells in order to successfully manage the wide variability bone loss scenarios seen in humans. The goals for this proposal are to assess the clinical potential of a "same day" ex vivo approach that can be easily adapted for human use; understand the role of transduced cells and host cells in bone formation and repair and; evaluate the safety of an ex vivo strategy using a lentiviral vector. The research proposed in Specific Aim 1 will compare the efficacy of a "same day" ev vivo strategy with the standard two step cell harvest and expansion approach. In Specific Aim 2 transgenic mice that express GFP under the control of cell differentiation specific promoters will be used to delineate the role of transduced cells and host cells in the healing of criticial sized bone defects. In Specific Aim 3 the safety of ex vivo gene therapy with a lentiviral vector will be assessed with a biodistribution assay and an evaluation of organ toxicity. These proposed Aims should provide valuable information regarding the feasibility, saftey, and biology of ex vivo gene therapy for bone repair. This data should move us one step closer to implementing this human use. PUBLIC HEALTH RELEVANCE: The ultimate goals of this research proposal are to enhance our knowledge of both the biology of bone repair and regional gene therapy, develop a new cost-effective treatment strategy and assess the safety of this therapy in order to move this technology closer to use in humans. In many cases our present treatment regimens are not adequate to manage difficult bone loss problems. Regional gene therapy has the potential to enhance our ability to treat difficult bone repair problems associated with fracture non-union, revision total joint arthroplasty and spinal fusion.

描述（由申请人提供）：离体区域基因疗法有可能解决与大量骨量缺乏、丰富的疤痕组织和有限的血管供应相关的骨质流失问题，目前还没有一致令人满意的解决方案。我们通过用腺病毒载体 (Ad-BMP-2) 或慢病毒载体 (LV-BMP-2) 转导的大鼠和人类骨细胞成功治愈临界大小的啮齿动物股骨缺损，证明了该策略的潜在临床实用性。含有 BMP-2 的 cDNA。最近对 Ad-BMP-2 与 LV-BMP-2 骨缺损愈合的比较进行了比较，使用 LV-BMP-2 治疗获得了质量更好的骨。我们推测，与慢病毒载体相关的 BMP 表达时间越长，慢病毒载体的效果越好，并且这种表达持续时间的增加可能是在患者所见的挑战性生物环境中实现充分骨修复所必需的。为了将基因疗法应用于临床，它必须具有成本效益、安全，并且必须更好地了解转导细胞骨修复的生物学，以便成功管理人类中广泛变化的骨质流失情况。该提案的目标是评估“同一天”离体方法的临床潜力，该方法可以轻松适应人类使用；了解转导细胞和宿主细胞在骨形成和修复中的作用；评估使用慢病毒载体的离体策略的安全性。具体目标 1 中提出的研究将比较“同一天”体外策略与标准两步细胞收获和扩增方法的功效。在Specific Aim 2转基因小鼠中，在细胞分化控制下表达GFP的特异性启动子将用于描述转导细胞和宿主细胞在修复临界大小的骨缺损中的作用。在具体目标 3 中，将通过生物分布测定和器官毒性评估来评估慢病毒载体离体基因治疗的安全性。这些提出的目标应提供有关骨修复离体基因治疗的可行性、安全性和生物学的有价值的信息。这些数据应该会让我们离实现这一人类用途更近一步。 公共健康相关性：该研究提案的最终目标是增强我们对骨修复生物学和区域基因治疗的了解，开发一种新的具有成本效益的治疗策略并评估该疗法的安全性，以使该技术更进一步用于人类。在许多情况下，我们目前的治疗方案不足以解决棘手的骨质流失问题。区域基因治疗有潜力提高我们治疗与骨折不愈合、翻修全关节置换术和脊柱融合相关的骨修复困难问题的能力。